Advertisement

Bacterial profile in human atherosclerotic plaques

  • Annika Lindskog Jonsson
    Affiliations
    The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden
    Search for articles by this author
  • Author Footnotes
    1 Present address: Food for Health Science Centre, Lund University, Lund, Sweden.
    Frida Fåk Hållenius
    Footnotes
    1 Present address: Food for Health Science Centre, Lund University, Lund, Sweden.
    Affiliations
    The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden
    Search for articles by this author
  • Rozita Akrami
    Affiliations
    The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden
    Search for articles by this author
  • Elias Johansson
    Affiliations
    Department of Public Health and Clinical Medicine, Umeå Stroke Centre, Umeå University, Umeå, Sweden

    Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden
    Search for articles by this author
  • Per Wester
    Affiliations
    Department of Public Health and Clinical Medicine, Umeå Stroke Centre, Umeå University, Umeå, Sweden

    Karolinska Institute Danderyds Hospital, Department of Clinical Sciences, Stockholm, Sweden
    Search for articles by this author
  • Conny Arnerlöv
    Affiliations
    Department of Surgical and Perioperative Sciences, Umeå University, Umeå, Sweden
    Search for articles by this author
  • Fredrik Bäckhed
    Correspondence
    Corresponding author. The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden.
    Affiliations
    The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden
    Search for articles by this author
  • Göran Bergström
    Correspondence
    Corresponding author. The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden.
    Affiliations
    The Wallenberg Laboratory, Department of Molecular and Clinical Medicine, University of Gothenburg, Bruna Stråket 16, 41345 Gothenburg, Sweden

    Department of Clinical Physiology, Sahlgrenska University Hospital, Gothenburg, Sweden
    Search for articles by this author
  • Author Footnotes
    1 Present address: Food for Health Science Centre, Lund University, Lund, Sweden.

      Highlights

      • Bacterial DNA is detected in human atherosclerotic plaques.
      • Bacterial composition is similar in plaques from asymptomatic and symptomatic patients.
      • Bacterial composition is similar in different plaque regions.
      • Plaque vulnerability is not explained by differences in bacterial DNA.

      Abstract

      Background and aims

      Several studies have confirmed the presence of bacterial DNA in atherosclerotic plaques, but its contribution to plaque stability and vulnerability is unclear. In this study, we investigated whether the bacterial plaque-profile differed between patients that were asymptomatic or symptomatic and whether there were local differences in the microbial composition within the plaque.

      Methods

      Plaques were removed by endarterectomy from asymptomatic and symptomatic patients and divided into three different regions known to show different histological vulnerability: A, upstream of the maximum stenosis; B, site for maximum stenosis; C, downstream of the maximum stenosis. Bacterial DNA composition in the plaques was determined by performing 454 pyrosequencing of the 16S rRNA genes, and total bacterial load was determined by qPCR.

      Results

      We confirmed the presence of bacterial DNA in the atherosclerotic plaque by qPCR analysis of the 16S rRNA gene but observed no difference (n.s.) in the amount between either asymptomatic and symptomatic patients or different plaque regions A, B and C. Unweighted UniFrac distance metric analysis revealed no distinct clustering of samples by patient group or plaque region. Operational taxonomic units (OTUs) from 5 different phyla were identified, with the majority of the OTUs belonging to Proteobacteria (48.3%) and Actinobacteria (40.2%). There was no difference between asymptomatic and symptomatic patients, or plaque regions, when analyzing the origin of DNA at phylum, family or OTU level (n.s.).

      Conclusions

      There were no major differences in bacterial DNA amount or microbial composition between plaques from asymptomatic and symptomatic patients or between different plaque regions, suggesting that other factors are more important in determining plaque vulnerability.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Moran A.E.
        • et al.
        Temporal trends in ischemic heart disease mortality in 21 world regions, 1980 to 2010: the Global Burden of Disease 2010 study.
        Circulation. 2014; 129: 1483-1492
        • Epstein S.E.
        • et al.
        Infection and atherosclerosis: potential roles of pathogen burden and molecular mimicry.
        Arterioscler. Thromb. Vasc. Biol. 2000; 20: 1417-1420
        • Libby P.
        • Egan D.
        • Skarlatos S.
        Roles of infectious agents in atherosclerosis and restenosis: an assessment of the evidence and need for future research.
        Circulation. 1997; 96: 4095-4103
        • Rosenfeld M.E.
        • Campbell L.A.
        Pathogens and atherosclerosis: update on the potential contribution of multiple infectious organisms to the pathogenesis of atherosclerosis.
        Thromb. Haemost. 2011; 106: 858-867
        • Ott S.J.
        • et al.
        Detection of diverse bacterial signatures in atherosclerotic lesions of patients with coronary heart disease.
        Circulation. 2006; 113: 929-937
        • Ziganshina E.E.
        • et al.
        Bacterial communities associated with atherosclerotic plaques from Russian individuals with atherosclerosis.
        PLoS One. 2016; 11: e0164836
        • Calandrini C.A.
        • et al.
        Microbial composition of atherosclerotic plaques.
        Oral Dis. 2014; 20: e128-e134
        • Koren O.
        • et al.
        Human oral, gut, and plaque microbiota in patients with atherosclerosis.
        Proc. Natl. Acad. Sci. U. S. A. 2011; 108: 4592-4598
        • Amar J.
        • et al.
        Involvement of tissue bacteria in the onset of diabetes in humans: evidence for a concept.
        Diabetologia. 2011; 54: 3055-3061
        • Amar J.
        • et al.
        Blood microbiota dysbiosis is associated with the onset of cardiovascular events in a large general population: the D.E.S.I.R. study.
        PLoS One. 2013; 8: e54461
        • Backhed F.
        • et al.
        The gut microbiota as an environmental factor that regulates fat storage.
        Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 15718-15723
        • Turnbaugh P.J.
        • et al.
        A core gut microbiome in obese and lean twins.
        Nature. 2009; 457: 480-484
        • Wellen K.E.
        • Hotamisligil G.S.
        Inflammation, stress, and diabetes.
        J. Clin. Invest. 2005; 115: 1111-1119
        • Karlsson F.H.
        • et al.
        Gut metagenome in European women with normal, impaired and diabetic glucose control.
        Nature. 2013; 498: 99-103
        • Yusuf S.
        • et al.
        Global burden of cardiovascular diseases: part I: general considerations, the epidemiologic transition, risk factors, and impact of urbanization.
        Circulation. 2001; 104: 2746-2753
        • Hansson G.K.
        Inflammation, atherosclerosis, and coronary artery disease.
        N. Engl. J. Med. 2005; 352: 1685-1695
        • Yilmaz A.
        • et al.
        Accumulation of immune cells and high expression of chemokines/chemokine receptors in the upstream shoulder of atherosclerotic carotid plaques.
        Exp. Mol. Pathol. 2007; 82: 245-255
        • Fagerberg B.
        • et al.
        Differences in lesion severity and cellular composition between in vivo assessed upstream and downstream sides of human symptomatic carotid atherosclerotic plaques.
        J. Vasc. Res. 2010; 47: 221-230
        • Salonen A.
        • et al.
        Comparative analysis of fecal DNA extraction methods with phylogenetic microarray: effective recovery of bacterial and archaeal DNA using mechanical cell lysis.
        J. Microbiol. Methods. 2010; 81: 127-134
        • Suzuki M.T.
        • Taylor L.T.
        • DeLong E.F.
        Quantitative analysis of small-subunit rRNA genes in mixed microbial populations via 5'-nuclease assays.
        Appl. Environ. Microbiol. 2000; 66: 4605-4614
        • Fierer N.
        • et al.
        The influence of sex, handedness, and washing on the diversity of hand surface bacteria.
        Proc. Natl. Acad. Sci. U. S. A. 2008; 105: 17994-17999
        • Caporaso J.G.
        • et al.
        QIIME allows analysis of high-throughput community sequencing data.
        Nat. Methods. 2010; 7: 335-336
        • Edgar R.C.
        Search and clustering orders of magnitude faster than BLAST.
        Bioinformatics. 2010; 26: 2460-2461
        • DeSantis T.Z.
        • et al.
        Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB.
        Appl. Environ. Microbiol. 2006; 72: 5069-5072
        • Wang Q.
        • et al.
        Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy.
        Appl. Environ. Microbiol. 2007; 73: 5261-5267
        • Caporaso J.G.
        • et al.
        PyNAST: a flexible tool for aligning sequences to a template alignment.
        Bioinformatics. 2010; 26: 266-267
        • Price M.N.
        • Dehal P.S.
        • Arkin A.P.
        FastTree 2–approximately maximum-likelihood trees for large alignments.
        PLoS One. 2010; 5: e9490
        • Faith D.P.
        Conservation evaluation and phylogenetic diversity.
        Biol. Conserv. 1992; 61: 1-10
        • Lozupone C.
        • Knight R.
        UniFrac: a new phylogenetic method for comparing microbial communities.
        Appl. Environ. Microbiol. 2005; 71: 8228-8235
        • Vazquez-Baeza Y.
        • et al.
        EMPeror: a tool for visualizing high-throughput microbial community data.
        Gigascience. 2013; 2: 16
        • Haas B.J.
        • et al.
        Chimeric 16S rRNA sequence formation and detection in Sanger and 454-pyrosequenced PCR amplicons.
        Genome Res. 2011; 21: 494-504
        • Olson F.J.
        • et al.
        Consistent differences in protein distribution along the longitudinal axis in symptomatic carotid atherosclerotic plaques.
        Biochem. Biophys. Res. Commun. 2010; 401: 574-580
        • Mitra S.
        • et al.
        In silico analyses of metagenomes from human atherosclerotic plaque samples.
        Microbiome. 2015; 3: 38
        • Havlik R.J.
        • et al.
        Unlikely association between clinically apparent herpesvirus infection and coronary incidence at older ages.
        Fram. Heart Study. Arteriosclerosis. 1989; 9: 877-880
        • Adler S.P.
        • et al.
        Prior infection with cytomegalovirus is not a major risk factor for angiographically demonstrated coronary artery atherosclerosis.
        J. Infect. Dis. 1998; 177: 209-212
        • Weiss S.M.
        • et al.
        Failure to detect Chlamydia pneumoniae in coronary atheromas of patients undergoing atherectomy.
        J. Infect. Dis. 1996; 173: 957-962
        • O'Connor C.M.
        • et al.
        Azithromycin for the secondary prevention of coronary heart disease events: the WIZARD study: a randomized controlled trial.
        JAMA. 2003; 290: 1459-1466
        • Grayston J.T.
        • et al.
        Azithromycin for the secondary prevention of coronary events.
        N. Engl. J. Med. 2005; 352: 1637-1645
        • Cannon C.P.
        • et al.
        Antibiotic treatment of Chlamydia pneumoniae after acute coronary syndrome.
        N. Engl. J. Med. 2005; 352: 1646-1654
        • Jespersen C.M.
        • et al.
        Randomised placebo controlled multicentre trial to assess short term clarithromycin for patients with stable coronary heart disease: CLARICOR trial.
        BMJ. 2006; 332: 22-27
        • Elkind M.S.
        • et al.
        Infectious burden and carotid plaque thickness: the northern Manhattan study.
        Stroke. 2010; 41: e117-e122
        • Georges J.L.
        • et al.
        Impact of pathogen burden in patients with coronary artery disease in relation to systemic inflammation and variation in genes encoding cytokines.
        Am. J. Cardiol. 2003; 92: 515-521
        • Zhu J.
        • et al.
        Effects of total pathogen burden on coronary artery disease risk and C-reactive protein levels.
        Am. J. Cardiol. 2000; 85: 140-146
        • Karlsson F.H.
        • et al.
        Symptomatic atherosclerosis is associated with an altered gut metagenome.
        Nat. Commun. 2012; 3: 1245
        • Winek K.
        • et al.
        Depletion of cultivatable gut microbiota by broad-spectrum antibiotic pretreatment worsens outcome after murine stroke.
        Stroke. 2016; 47: 1354-1363
        • Benakis C.
        • et al.
        Commensal microbiota affects ischemic stroke outcome by regulating intestinal gammadelta T cells.
        Nat. Med. 2016; 22: 516-523
        • Olson F.J.
        • et al.
        Increased vascularization of shoulder regions of carotid atherosclerotic plaques from patients with diabetes.
        J. Vasc. Surg. 2011; 54 (e5): 1324-1331